PROJECT SUMMARY Campylobacter infection is a leading cause of human foodborne illness in the United States. Although human campylobacteriosis is usually associated with the consumption of contaminated food, pets can be another source of infection. Dogs and cats are not commonly thought of as a source for human infection, but Campylobacter carriage is well-recognized in these pets. Risk factors for Campylobacter infection in dogs and cats include consumption of raw meat diets and contaminated treats such as uncooked beef bones. People can be exposed to Campylobacter when handling contaminated pet food and treats or feces from infected pets. Despite recognition of these cases and the potential risk of campylobacteriosis in dogs and cats, Campylobacter culture is not routinely performed in these species. Members of the Texas A&M University Veterinary Medical Teaching Hospital Clinical Microbiology Laboratory (TAMU CML), a FDA Vet-LIRN laboratory, have previously tested primers for Campylobacter jejuni that target cpn60 for detection of C. jejuni from canine fecal samples. Recent work by the Ohio Vet-LIRN laboratory indicates that primers that target gyr provide improved detection of C. jejuni in canine feces. The goal of this proposal is to validate molecular methods to detect Campylobacter shedding in dogs and cats because 1) Campylobacter infection is well-recognized in dogs and cats; 2) dogs and cats live in close association with people, particularly children; 3) canine and feline shedding of Campylobacter spp. is well-recognized and associated with human infection; 4) dogs and cats are not routinely screened for Campylobacter shedding despite recognition of the risk of Campylobacter infection in dogs and cats; and 5) the absence of standardized methods for Campylobacter detection make it difficult to compare studies from different geographic areas and different laboratories. Members of the TAMU CML will validate molecular methods to detect C. jejuni gyrA in canine feces and test these methods in 5 independent, blinded laboratories with C. jejuni spiked test samples. Additionally, the collaborative effort of test validation and sample exchange between the 5 collaborating laboratories will strengthen the ties between these Vet-LIRN laboratories thereby promoting seamless interactions during actual emergency related events.